目的：探讨碱性亮氨酸拉链转录因子ATF样蛋白3（BATF3）在肾透明细胞癌（ccRCC）组织中的表达及其调控ccRCC细胞恶性生物学行为的分子机制。方法：收集2019 年3月至2022 年1月间在中国人民解放军联勤保障部队第九八〇医院手术切除的64例ccRCC组织和相应癌旁组织，qPCR 法检测ccRCC组织、癌旁组织和肾癌ACHN、786-O细胞中BATF3 mRNA的表达，免疫组化检测ccRCC 组织、癌旁组织中BATF3蛋白的表达，并分析其与临床病理特征的关系。构建BATF3敲减及过表达质粒，分别转染786-O、ACHN 细胞，通过MTS 法、Transwell 实验检测BATF3 对786-O 或ACHN 细胞增殖、迁移、侵袭能力的影响，qPCR 法检测敲减或过表达BATF3 对786-O 或ACHN 细胞EMT 相关基因表达的影响，CHIP、双荧光素酶报告基因实验检测BATF3是否与波形蛋白（VIM）启动子区结合并调控其转录，MTS法、Transwell 实验检测同时过表达BATF3及敲减VIM对786-O细胞增殖、迁移、侵袭能力的影响。结果：与癌旁组织比较，ccRCC 组织中BATF3的mRNA和蛋白均呈高表达(均P<0.01)，并且BATF3 mRNA 与ccRCC 的分化程度和TNM分期密切关联（均P<0.01）；与正常肾上皮细胞293T相比，BATF3 在ACHN 及786-O细胞中均呈高表达（均P<0.01）。敲减BATF3 表达均能明显抑制786-O 细胞的增殖、迁移、侵袭能力(均P<0.01)，过表达BATF3则均能促进ACHN细胞的增殖、迁移和侵袭能力（均P<0.01），敲减或过表达BATF3能抑制786-O 细胞或促进ACHN细胞的EMT相关基因的表达（均P<0.01）。BATF3可与VIM启动子区的位点结合，直接调控VIM的转录表达。同时过表达BATF3及敲减VIM可逆转过表达BATF3对786-O 细胞增殖、迁移、侵袭能力的影响。结论：BATF3在ccRCC 组织中呈高表达，并与其分化程度和TNM 分期密切关联；BATF3 通过调控VIM 的表达影响ACHN、786-O 细胞的恶性生物学行为，其可作为临床治疗ccRCC的潜在靶点。

Objective: To investigate the expression of basic leucine zipper ATF-like transcription factor 3 (BATF3) in clear cell renal cell carcinoma (ccRCC) and the molecular mechanism of its regulation of malignant biological behavior of ccRCC cells. Methods: ccRCC tissues and para-cancerous tissues were collected from 64 ccRCC patients who underwent surgical treatment at the 980th Hospital of the Joint Logistics Support Force of PLA from March 2019 to January 2022. The expression of BATF3 mRNA in ccRCC tissues, para-cancerous tissues, renal carcinoma ACHN and 786-O cells was determined by qPCR. The expressions of BATF3 protein in ccRCC tissues and para-cancerous tissues were determined by immunohistochemistry and the relationship between the expressions and the clinicopathologic characteristics was analyzed. BATF3 knockdown and over-expression plasmids were constructed and transfected into 786-O and ACHN cells, and the effects of BATF3 on the proliferation, migration and invasion of 786-O cells or ACHN cells were determined by MTS and Transwell assays. The effects of BATF3 on the expressions of related epithelial-mesenchymal transition (EMT) genes of 786-O and ACHN cells were detected by qPCR. CHIP and dual luciferase reporter assay were performed to detect the binding of BATF3 as a transcription factor to vimentin (VIM) and regulate its transcription. The effects of simultaneous over-expression of BATF3 and knockdown of VIM on the proliferation, migration and invasion ability of 786-O cells were determined by MTS and Transwell assays. Results: Compared with para-cancerous tissues, BATF3 mRNA and protein expressions were markedly higher in ccRCC tissues (all P<0.01) and the expression level of BATF3 mRNA was closely correlated with the degree of differentiation of ccRCC and its TNM stage (all P<0.01). Compared with normal renal epithelial cells 293T, the expression of BATF3 was significantly higher in ccRCC cells ACHN and 786-O (all P<0.01). Knockdown of BATF3 expression significantly inhibited the proliferation,migration and invasion ability of 786-O cells (all P<0.01). Over-expression of BATF3 significantly promoted the proliferation,migration and invasion ability of ACHN cells (all P<0.01). Knockdown or overexpression of BATF3 inhibited the expression of EMT-related genes in 786-O cells or promoted the expression of EMT-related genes in ACHN cells (all P<0.01). BATF3 could bind to sites in the upstream promoter region of VIM and directly regulate the transcription expression of VIM. Simultaneous overexpression of BATF3 and knockdown of VIM reversed the effects of over-expression of BATF3 on the proliferation, migration and invasion ability of 786-O cells. Conclusion: BATF3 was highly expressed in ccRCC tissues and was closely related to its differentiation degree and TNM stage. BATF3 directly regulated the expression of VIM, which in turn regulated the malignant biological behavior of ACHN and 786-O cells. Therefore, it could be used as a potential target for clinical treatment of ccRCC.

第九八〇医院科学技术孵育计划（No. FYJHZD202302，No. FYJHMS202312）